Funds are requested for purchase of the Amersham-Pharmacia Fluorescence 2D Differential Gel Electrophoresis system, consisting of Typhoon 9400 Scanner, Decyder software package for image analysis, associated Cy-dyes for multiplex fluorescence analysis, and a single high capacity electrophoresis system for running 12D gels. This system will be utilized by three groups of NIH-funded investigators whose projects either employ proteomics technology currently or would be materially strengthened by its addition. The research programs to be enhanced by acquisition of this package have direct connections to microbial pathogenesis, cardiovascular disease, cancer, and development and differentiation in eukaryotes. An administrative structure and operating policy is described, and evidence of both institutional commitment to operation and support and a financial plan for long-term maintenance is provided. Major and secondary user groups containing 6 and 12 investigators respectively are anticipated to account for approximately 85% of available instrument time, leaving approximately 15% for other investigators and/or new research initiatives. Acquisition of the 2D DIGE system will materially enhance a wide variety of research programs aimed at understanding qualitative and quantitative changes in cellular proteomes in response to infections, disease states, and environmental cues.